+Open data
-Basic information
Entry | Database: PDB / ID: 7jr9 | ||||||||||||
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Title | Chlamydomonas reinhardtii radial spoke minimal head complex | ||||||||||||
Components |
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Keywords | STRUCTURAL PROTEIN / Cilia / Radial spoke / Mechano-regulation | ||||||||||||
Function / homology | Function and homology information radial spoke head / radial spoke / kinocilium / cilium movement involved in cell motility / motile cilium assembly / axoneme assembly / motile cilium / axoneme / membrane Similarity search - Function | ||||||||||||
Biological species | Chlamydomonas reinhardtii (plant) | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.95 Å | ||||||||||||
Authors | Grossman-Haham, I. / Coudray, N. / Yu, Z. / Wang, F. / Bhabha, G. / Vale, R.D. | ||||||||||||
Funding support | United States, 3items
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Citation | Journal: Nat Struct Mol Biol / Year: 2021 Title: Structure of the radial spoke head and insights into its role in mechanoregulation of ciliary beating. Authors: Iris Grossman-Haham / Nicolas Coudray / Zanlin Yu / Feng Wang / Nan Zhang / Gira Bhabha / Ronald D Vale / Abstract: Motile cilia power cell locomotion and drive extracellular fluid flow by propagating bending waves from their base to tip. The coordinated bending of cilia requires mechanoregulation by the radial ...Motile cilia power cell locomotion and drive extracellular fluid flow by propagating bending waves from their base to tip. The coordinated bending of cilia requires mechanoregulation by the radial spoke (RS) protein complexes and the microtubule central pair (CP). Despite their importance for ciliary motility across eukaryotes, the molecular function of the RSs is unknown. Here, we reconstituted the Chlamydomonas reinhardtii RS head that abuts the CP and determined its structure using single-particle cryo-EM to 3.1-Å resolution, revealing a flat, negatively charged surface supported by a rigid core of tightly intertwined proteins. Mutations in this core, corresponding to those involved in human ciliopathies, compromised the stability of the recombinant complex, providing a molecular basis for disease. Partially reversing the negative charge on the RS surface impaired motility in C. reinhardtii. We propose that the RS-head architecture is well-suited for mechanoregulation of ciliary beating through physical collisions with the CP. | ||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 7jr9.cif.gz | 233.2 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7jr9.ent.gz | 188.1 KB | Display | PDB format |
PDBx/mmJSON format | 7jr9.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7jr9_validation.pdf.gz | 758.8 KB | Display | wwPDB validaton report |
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Full document | 7jr9_full_validation.pdf.gz | 764 KB | Display | |
Data in XML | 7jr9_validation.xml.gz | 37.1 KB | Display | |
Data in CIF | 7jr9_validation.cif.gz | 58.6 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/jr/7jr9 ftp://data.pdbj.org/pub/pdb/validation_reports/jr/7jr9 | HTTPS FTP |
-Related structure data
Related structure data | 22444MC 7jrjC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
-Radial spoke protein ... , 2 types, 3 molecules ABE
#1: Protein | Mass: 29572.348 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Chlamydomonas reinhardtii (plant) / Gene: RSP9, CHLRE_07g330200v5, CHLREDRAFT_182960 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q27YU5 #6: Protein | | Mass: 23553.943 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Chlamydomonas reinhardtii (plant) / Gene: RSP10, CHLRE_01g005450v5, CHLREDRAFT_185792 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q27YU4 |
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-Flagellar radial spoke protein ... , 4 types, 4 molecules CDFG
#2: Protein | Mass: 52159.195 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Chlamydomonas reinhardtii (plant) / Gene: RSP4 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q01656 |
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#3: Protein | Mass: 48884.551 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Chlamydomonas reinhardtii (plant) / Gene: RSP6 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q01657 |
#4: Protein/peptide | Mass: 698.854 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Chlamydomonas reinhardtii (plant) / Production host: Spodoptera frugiperda (fall armyworm) |
#5: Protein/peptide | Mass: 528.644 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Chlamydomonas reinhardtii (plant) / Production host: Spodoptera frugiperda (fall armyworm) |
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: radial spoke minimal head complex / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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Molecular weight | Experimental value: NO |
Source (natural) | Organism: Chlamydomonas reinhardtii (plant) |
Source (recombinant) | Organism: Spodoptera frugiperda (fall armyworm) |
Buffer solution | pH: 7.4 / Details: 30 mM HEPES, 150 mM NaCl, 1 mM TCEP |
Specimen | Conc.: 0.1 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Grid material: GOLD / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 95 % / Chamber temperature: 293 K |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 130000 X / Nominal defocus max: 2500 nm / Nominal defocus min: 1000 nm / Cs: 0.001 mm |
Image recording | Electron dose: 73.3 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 2 / Num. of real images: 2886 |
EM imaging optics | Energyfilter slit width: 30 eV |
Image scans | Movie frames/image: 35 |
-Processing
Software | Name: PHENIX / Version: 1.17.1_3660: / Classification: refinement | ||||||||||||||||||||||||||||||||||||
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EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 1775590 | ||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 2.95 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 251088 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||
Atomic model building | Protocol: AB INITIO MODEL | ||||||||||||||||||||||||||||||||||||
Refinement | Cross valid method: NONE |